Firefly and Renilla luciferases are widely used as reporter genes for research and drug screening. Our luciferase kits offer excellent sensitivity at a fraction of the cost of other suppliers. Our lyophilized firefly assay kits can be shipped at room temperature to save you even more money. We also offer a wide selection of D-Luciferin and coelenterazine substrates available separately.
Bioluminenscent Enzyme Reactions
Most cells have no intrinsic bioluminescence or luciferase activity, making luciferase reporter assays highly specific and sensitive. The bioluminescent signal of firefly luciferase is generated by the ATP-catalyzed oxidation of D-luciferin into oxyluciferin, which releases light. This is a highly efficient reaction, which makes this system extremely sensitive and linear as a reporter assay. The light emitted by the reaction can be measured by a luminometer, and signal strength can be directly correlated to the amount of firefly luciferase protein expressed.
Renilla luciferase enzyme from sea pansy catalyzes the oxidation of coelenterazine to generate light. Because firefly and Renilla luciferases use different substrates, they can be used in multiplex assays, typically with one enzyme serving as normalization control for transfection efficiency, and the other enzyme serving as a reporter enzyme.
Reporter Gene Assays
Several commercially luciferase reporter assay kits are available that contain an expression vector containing the luciferase gene. A luciferase reporter assay is usually performed to investigate the strength and regulation of a promoter. This assay has three general steps: 1) clone your promoter of interest upstream of the luciferase gene in an expression vector; 2) transfect cells with the expression vector, and allow cells to grow to express the luciferase; 3) lyse the cells and measure luciferase expression with a luminometer. For this step we recommend our kits described below, which provide lysis reagents, luciferin and buffers optimized for high sensitivity and linearity.
Cell Viability Assays
Bioluminescent assays can be used to monitor cell viability by quantitating ATP. Because ATP is an indicator of metabolically active cells, the number of viable cells can be assessed based on the amount of ATP available.